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Mycoplasma detection – biochemical test

Mycoplasma detection – biochemical test

The test is intended to detect mycoplasma in cell culture media. The test uses selective evidence of activity of those mycoplasma enymes present in the vast majority of the 180 known species of mycoplasma, however not present in eucaryotic cells.

Principle of the test

The presence of mycoplasma in a cell culture is highly undesirable and can conclude in a serious distortion or even failure of in vitro experimental results coming from:

  • undesirable changes of cell morphology
  • inhibition of cell metabolism
  • alteration of cell membrane transport
  • inhibition of cell growth
  • alteration of DNA and chromosomal aberrations

A selective biochemical test exploits the activity of mycoplasma enzymes present in most of 180 mycoplasma species, but not occurring in eukaryotic cells. If the tested sample is mycoplasma positive, the supernatant after cell lysis contains mycoplasma enzymes that change ADP to ATP. In a subsequent step, the ATP acts as a substrate for luciferase which results in emitting luminescence. Contamination is excluded / evidenced by measuring the ratio of luminescence prior to and after luciferase addition.

Parameters of the test

Test detection limit: 50 cfu/ml

Requirements for the material to be tested

Customer provides us with 1 to 2 ml of cultivation medium, made according to instructions below.

Preparing cells to be tested

To ensure reliable test results, please keep carefully to the recommendations as follows:

  • After defrosting the cell stock from liquid nitrogen, cultivate the cell culture in standard cultivation conditions for at least 14 days.
  • Cultivate the cells in the amount of medium recommended for a given cultivation container, do not exceed the recommended amount.
  • Cell confluency must be at least 80 % at the moment of medium withdrawal.
  • Withdrawal of a medium sample to be tested must be performed no sooner than 2 days following the last exchange of medium.
  • Centrifuge 1-2 ml of withdrawn cultivation medium (200 x g, 5 minutes, at room temperature) and transfer into a new tube (sufficient volume is 500 μl).
  • The medium can be stored up to 6 months at -80 °C.

Sample delivery

if the delivery time does not exceed 24 hours, send samples chilled to 2 to 8 °C (on wet ice), otherwise we recommend delivery on dry ice.

Output of the test

We provide you with a protocol in English with a statement if the presence of mycoplasma was detected.

Cat. No. Product   Price
1830 Mycoplasma detection - biochemical test
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